Synaptic vesicle pool size, release probability and synaptic depression are sensitive to Ca2+ buffering capacity in the developing rat calyx of Held

The calyx of Held, a specialized synaptic terminal in the medial nucleus of the trapezoid body, undergoes a series of changes during postnatal development that prepares this synapse for reliable high frequency firing. These changes reduce short-term synaptic depression during tetanic stimulation and thereby prevent action potential failures during a stimulus train. We measured presynaptic membrane capacitance changes in calyces from young postnatal day 5-7 (p5-7) or older (p10-12) rat pups to examine the effect of calcium buffer capacity on vesicle pool size and the efficiency of exocytosis. Vesicle pool size was sensitive to the choice and concentration of exogenous Ca2+ buffer, and this sensitivity was much stronger in younger animals. Pool size and exocytosis efficiency in p5-7 calyces were depressed by 0.2 mM EGTA to a greater extent than with 0.05 mM BAPTA, even though BAPTA is a 100-fold faster Ca2+ buffer. However, this was not the case for p10-12 calyces. With 5 mM EGTA, exocytosis efficiency was reduced to a much larger extent in young calyces compared to older calyces. Depression of exocytosis using pairs of 10-ms depolarizations was reduced by 0.2 mM EGTA compared to 0.05 mM BAPTA to a similar extent in both age groups. These results indicate a developmentally regulated heterogeneity in the sensitivity of different vesicle pools to Ca2+ buffer capacity. We propose that, during development, a population of vesicles that are tightly coupled to Ca2+ channels expands at the expense of vesicles more distant from Ca2+ channels.

Liver lipid peroxidation and antioxidant capacity in cerulein-induced acute pancreatitis

The aim of this study was to evaluate the role of oxidative damage in pancreatitis-induced hepatic injury. Thirty-five rats were divided into five groups (each of 7 rats): control, cerulein (100 µg/kg body weight), cerulein and pentoxifylline (12 mg/kg body weight), cerulein plus L-NAME (10 mg/kg body weight) and cerulein plus L-arginine (160 mg/kg body weight). The degree of hepatic cell degeneration differed significantly between groups. Mean malondialdehyde levels were 7.00 ± 2.29, 20.89 ± 10.13, 11.52 ± 4.60, 18.69 ± 8.56, and 8.58 ± 3.68 nmol/mg protein for the control, cerulein, pentoxifylline, L-NAME, and L-arginine groups, respectively. Mean catalase activity was 3.20 ± 0.83, 1.09 ± 0.35, 2.05 ± 0.91, 1.70 ± 0.60, and 2.85 ± 0.47 U/mg protein for the control, cerulein, pentoxifylline, L-NAME, and L-arginine groups, respectively, and mean glutathione peroxidase activity was 0.72 ± 0.25, 0.33 ± 0.09, 0.37 ± 0.04, 0.34 ± 0.07 and 0.42 ± 0.1 U/mg protein for the control, cerulein, pentoxifylline, L-NAME, and L-arginine groups, respectively. Cerulein-induced liver damage was accompanied by a significant increase in tissue malondialdehyde levels (P < 0.05) and a significant decrease in catalase (P < 0.05) and GPx activities (P < 0.05). L-arginine and pentoxifylline, but not L-NAME, protected against this damage. Oxidative injury plays an important role not only in the pathogenesis of AP but also in pancreatitis-induced hepatic damage.

A link between sleep loss, glucose metabolism and adipokines

The present review evaluates the role of sleep and its alteration in triggering problems of glucose metabolism and the possible involvement of adipokines in this process. A reduction in the amount of time spent sleeping has become an endemic condition in modern society, and a search of the current literature has found important associations between sleep loss and alterations of nutritional and metabolic contexts. Studies suggest that sleep loss is associated with problems in glucose metabolism and a higher risk for the development of insulin resistance and type 2 diabetes mellitus. The mechanism involved may be associated with the decreased efficacy of regulation of the hypothalamus-pituitary-adrenal axis by negative feedback mechanisms in sleep-deprivation conditions. In addition, changes in the circadian pattern of growth hormone (GH) secretion might also contribute to the alterations in glucose regulation observed during sleep loss. On the other hand, sleep deprivation stress affects adipokines - increasing tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) and decreasing leptin and adiponectin -, thus establishing a possible association between sleep-debt, adipokines and glucose metabolism. Thus, a modified release of adipokines resulting from sleep deprivation could lead to a chronic sub-inflammatory state that could play a central role in the development of insulin resistance and type 2 diabetes mellitus. Further studies are necessary to investigate the role of sleep loss in adipokine release and its relationship with glucose metabolism.

In vitro proliferation and differentiation of hepatic oval cells and their potential capacity for intrahepatic transplantation

Hepatic oval cells (HOCs) are recognized as facultative liver progenitor cells that play a role in liver regeneration after acute liver injury. Here, we investigated the in vitro proliferation and differentiation characteristics of HOCs in order to explore their potential capacity for intrahepatic transplantation. Clusters or scattered HOCs were detected in the portal area and interlobular bile duct in the liver of rats subjected to the modified 2-acetylaminofluorene and partial hepatectomy method. Isolated HOCs were positive for c-kit and CD90 staining (99.8% and 88.8%, respectively), and negative for CD34 staining (3.6%) as shown by immunostaining and flow cytometric analysis. In addition, HOCs could be differentiated into hepatocytes and bile duct epithelial cells after leukemia inhibitory factor deprivation. A two-cuff technique was used for orthotopic liver transplantation, and HOCs were subsequently transplanted into recipients. Biochemical indicators of liver function were assessed 4 weeks after transplantation. HOC transplantation significantly prolonged the median survival time and improved the liver function of rats receiving HOCs compared to controls (P=0.003, Studentt-test). Administration of HOCs to rats also receiving liver transplantation significantly reduced acute allograft rejection compared to control liver transplant rats 3 weeks following transplantation (rejection activity index score: control=6.3±0.9; HOC=3.5±1.5; P=0.005). These results indicate that HOCs may be useful in therapeutic liver regeneration after orthotopic liver transplantation.

Diastolic function is associated with quality of life and exercise capacity in stable heart failure patients with reduced ejection fraction

Exercise capacity and quality of life (QOL) are important outcome predictors in patients with systolic heart failure (HF), independent of left ventricular (LV) ejection fraction (LVEF). LV diastolic function has been shown to be a better predictor of aerobic exercise capacity in patients with systolic dysfunction and a New York Heart Association (NYHA) classification ≥II. We hypothesized that the currently used index of diastolic function E/e' is associated with exercise capacity and QOL, even in optimally treated HF patients with reduced LVEF. This prospective study included 44 consecutive patients aged 55±11 years (27 men and 17 women), with LVEF<0.50 and NYHA functional class I-III, receiving optimal pharmacological treatment and in a stable clinical condition, as shown by the absence of dyspnea exacerbation for at least 3 months. All patients had conventional transthoracic echocardiography and answered the Minnesota Living with HF Questionnaire, followed by the 6-min walk test (6MWT). In a multivariable model with 6MWT as the dependent variable, age and E/e' explained 27% of the walked distance in 6MWT (P=0.002; multivariate regression analysis). No association was found between walk distance and LVEF or mitral annulus systolic velocity. Only normalized left atrium volume, a sensitive index of diastolic function, was associated with decreased QOL. Despite the small number of patients included, this study offers evidence that diastolic function is associated with physical capacity and QOL and should be considered along with ejection fraction in patients with compensated systolic HF.

Lipid peroxidation, detoxification capacity, and genome damage in mice after transplacental exposure to pharmaceutical drugs

Data on genome damage, lipid peroxidation, and levels of glutathione peroxidase (GPX) in newborns after transplacental exposure to xenobiotics are rare and insufficient for risk assessment. The aim of the current study was to analyze, in an animal model, transplacental genotoxicity, lipid peroxidation, and detoxification disturbances caused by the following drugs commonly prescribed to pregnant women: paracetamol, fluconazole, 5-nitrofurantoin, and sodium valproate. Genome damage in dams and their newborn pups transplacentally exposed to these drugs was investigated using the in vivo micronucleus (MN) assay. The drugs were administered to dams intraperitoneally in three consecutive daily doses between days 12 and 14 of pregnancy. The results were correlated, with detoxification capacity of the newborn pups measured by the levels of GPX in blood and lipid peroxidation in liver measured by malondialdehyde (HPLC-MDA) levels. Sodium valproate and 5-nitrofurantoin significantly increased MN frequency in pregnant dams. A significant increase in the MN frequency of newborn pups was detected for all drugs tested. This paper also provides reference levels of MDA in newborn pups, according to which all drugs tested significantly lowered MDA levels of newborn pups, while blood GPX activity dropped significantly only after exposure to paracetamol. The GPX reduction reflected systemic oxidative stress, which is known to occur with paracetamol treatment. The reduction of MDA in the liver is suggested to be an unspecific metabolic reaction to the drugs that express cytotoxic, in particular hepatotoxic, effects associated with oxidative stress and lipid peroxidation.

Plasticity of neutrophils reveals modulatory capacity

Neutrophils are widely known as proinflammatory cells associated with tissue damage and for their early arrival at sites of infection, where they exert their phagocytic activity, release their granule contents, and subsequently die. However, this view has been challenged by emerging evidence that neutrophils have other activities and are not so short-lived. Following activation, neutrophil effector functions include production and release of granule contents, reactive oxygen species (ROS), and neutrophil extracellular traps (NETs). Neutrophils have also been shown to produce a wide range of cytokines that have pro- or anti-inflammatory activity, adding a modulatory role for this cell, previously known as a suicide effector. The presence of cytokines almost always implies intercellular modulation, potentially unmasking interactions of neutrophils with other immune cells. In fact, neutrophils have been found to help B cells and to modulate dendritic cell (DC), macrophage, and T-cell activities. In this review, we describe some ways in which neutrophils influence the inflammatory environment in infection, cancer, and autoimmunity, regulating both innate and adaptive immune responses. These cells can switch phenotypes and exert functions beyond cytotoxicity against invading pathogens, extending the view of neutrophils beyond suicide effectors to include functions as regulatory and suppressor cells.

Resistência de quatro genótipos de amendoim à produção de aflatoxina B1 após inoculação com Aspergillus flavus Link

Avaliou-se a produção de aflatoxina B1 pelo Aspergillus flavus IMI 190443, inoculado em sementes de genótipos de amendoim: Tatu Vermelho, VRR-245, 2117 e 2155, in natura e autoclavado a 1210 C por 20 minutos, previamente cultivados no Centro Experimental do Instituto Agronômico de Campinas, em 1995/96. A aflatoxina B1 foi quantificada por cromatografia em camada delgada, utilizando comparação visual com padrões. Os níveis de aflatoxina B1 das amostras autoclavadas e in natura dos genótipos Tatu Vermelho, VRR-245 e 2155 foram significativamente diferentes (p <0,05, Teste de Duncan) quando comparados aos níveis do genótipo 2117. O genótipo 2117, originário da Índia, apresentou independente do processo com e sem aquecimento os menores níveis de aflatoxina B1 em relação aos dos outros genótipos. Os resultados obtidos mostram perspectivas de exploração da resistência varietal no controle da aflatoxina.

Composição, proteólise, capacidade de derretimento e formação de "blisters" do queijo mussarela obtido pelos métodos tradicional e de ultrafiltração: composition, proteolysis, melting capacity and blisters formation

O objetivo deste trabalho foi comparar a composição, a proteólise, a capacidade de derretimento e a formação de "blisters" (bolhas) em queijos tipo Mussarela fabricados com retentado de leite (MR) de fator de concentração volumétrica (FCV) de 2,34:1, com um queijo Mussarela padrão (MP) fabricado com leite não ultrafiltrado. Foi realizado um ensaio de produção com 3 lotes de MR e um lote de MP. Determinou-se a composição do leite, retentado, soro, água de filagem e queijos e a proteólise, a capacidade de derretimento e a formação de "blisters" nos queijos com 7, 15, 30 e 60 dias de armazenamento refrigerado. MRs apresentaram maiores valores de pH e de porcentagem de cinzas e de proteína total e menores porcentagens de acidez titulável, gordura, gordura no extrato seco e sal quando comparadas a MP. Durante o tempo de estocagem, as MRs apresentaram menor proteólise e capacidade de derretimento, em todas as datas analisadas. A porcentagem de área coberta por 'blisters" na pizza e o diâmetro médio dos mesmos foram maiores na MP durante o primeiro mês de estocagem e depois ambos os tipos de queijos apresentaram comportamentos similares para estes 2 parâmetros.

Evaluation of thermotolerant capacity of lactic acid bacteria isolated from commercial sausages and the effects of their addition on the quality of cooked sausages

The thermotolerant capacity of several lactic acid bacteria strains isolated from cooked commercial sausages was determined. Four strains were positively identified as Lactobacillus plantarum, Lactobacillus curvatus, Pediococcus pentosaceus and Pediococcus acidilacti, after surviving thermal treatment (70 °C during 60 minutes). Thermotolerant strains were inoculated in sausage batters before cooking in order to determine their effect on color, texture, acceptance and inhibition of Enterobacteria during 12 days at 8 °C. No significant effect of the inoculated strains was detected on color parameters. Textural profile parameters, cohesiveness and resilience, were not affected by the inoculation of thermotolerant lactic acid bacteria, but L. curvatus sausages resulted softer than the rest of the treatments. Samples inoculated with L. curvatus also obtained the lowest scores for the sensory attributes evaluated, with the remaining treatments causing no unfavorable effects on sausage acceptance. There was a reduction in enterobacterial counts after 12 days of cold storage in inoculated samples. The performance of inoculated lactic acid bacteria strains can be explained in a similar way as that of starter cultures in dry-fermented sausages, where the growth in nests impairs other pathogenic microorganisms present in the rest of the sausage, since environmental conditions and the early inoculation of these thermotolerant strains favor them to become the dominant flora.

Comparison of total phenolic content and antiradical capacity of powders and "chocolates" from cocoa and cupuassu

Polyphenolic compounds seem to be related to the health benefits produced by the cocoa due to their antioxidant properties. Cupuassu powder, prepared from Theobroma grandiflorum seeds, is a very promising cocoa powder substitute. In order to assess the potential health benefits of the cupuassu powder, a comparison was performed between cocoa, chocolate, and cupuassu powders in relation to the content of total phenolic compounds, flavonoids and DPPH scavenging capacity of methanolic extracts. Cupuassu "chocolates" (milk, dark, and white) were also analyzed. Mineral, lipid, protein, and moisture determinations were made in cocoa and cupuassu powders and in cupuassu "chocolates". Results showed that the phenolic contents of cocoa and chocolate powders are more than three times higher than those of cupuassu powder; however, flavonoid contents were significantly lower. The DPPH scavenging capacity varied hugely among the different samples, from 0.5 (white cupuassu "chocolate") to 120 (cocoa powder) μg of Trolox equivalent per 100 g (FW), and presented a significant correlation (r = 0.977) with the total phenolic contents but not with the flavonoid contents (r = -0.035).

Flavonoids, total phenolics and antioxidant capacity: comparison between commercial green tea preparations

The potential health benefits attributed to green tea and its catechins such as antioxidant effects, cancer chemoprevention, and weight loss have led to a huge increase of green tea products in the food market. The objectives of this work were to analyze and compare these products in terms of phenolic contents and in vitro antioxidant capacity including tea bags, dehydrated leaves, and ready-to-drink preparations after standardization of the infusion preparation procedure. Total phenolics content in 1 cup of the different teas varied from 90 to 341 mg of catechin equivalents, and the highest and the lowest values were both those of the ready-to-drink products. Infusions prepared from tea bags had contents varying from 96 to 201 mg.200 mL-1, and there were no significant differences among batches. The DPPH radical scavenging and the Oxygen Radical Absorbing Capacities (ORAC) varied largely among the different tea preparations, from 23 to 131 mmoles of Trolox Equivalents (TE).200 mL-1 (DPPH), and from 1.2 to 5.1 mmoles of TE.200 mL-1 (ORAC), but again there were no differences among infusions or ready-to-drink commercial preparations. However, the antioxidant capacity of ready-to-drink products was partially due to the presence of other non-phenolic compounds such as ascorbic acid

Comparison of phenol content and antioxidant capacity of nuts

Frequent nut intake is associated with protective effects against cardiovascular diseases. In addition to the generally high contents of unsaturated fatty acids, polyphenol compounds seem to be also implicated in health promoting effects of nuts due to their antioxidant properties. In this way, eleven different kinds of nuts, including pinhao seeds (Araucaria angustifolia) and Brazil nuts (Bertholletia excelsa), typical of Brazil, were analyzed for the content of phenol compounds, including the potent anti-mutagenic and anti-cancer ellagic acid, and antioxidant capacity of methanolic extracts. The antioxidant capacity varied a hundred times among the different nuts, from 1.2 to 120 mg of Trolox equivalents.100 g-1 (FW). Total ellagic acid was determined after acid hydrolysis of ellagitannins and ellagic acid glycosides, and it was detected in only 3 of the 11 samples. The content of free and total ellagic acid in nuts varied from 0.37 to 41 and from 149 (chestnuts) to 823 (walnuts) mg.100 g-1 (FW), respectively. Among nuts, samples with the highest contents of ellagic acid (walnuts and pecans) also presented the highest total phenol contents and DPPH radical scavenging capacities. Pinhao seeds and Brazil nuts did not present significant amounts of phenols nor antioxidant capacity.

Antioxidant capacity of anthocyanins from acerola genotypes

Anthocyanins from 12 acerola genotypes cultivated at the Active Germplasm Bank at Federal Rural University of Pernambuco were isolated for antioxidant potential evaluation. The antioxidant activity and radical scavenging capacity of the anthocyanin isolates were measured according to the β-carotene bleaching method and 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical scavenging assay, respectively. The antioxidant activity varied from 25.58 to 47.04% at 0.2 mg.mL-1, and it was measured using the β-carotene bleaching method. The free radical scavenging capacity increased according to the increase in concentration and reaction time by the DPPH assay. At 16.7 μg.mL-1 concentration and after 5 minutes and 2 hours reaction time, the percentage of scavenged radicals varied from 36.97 to 63.92% and 73.27 to 94.54%, respectively. Therefore, the antioxidant capacity of acerola anthocyanins varied amongst acerola genotypes and methods used. The anthocyanins present in this fruit may supply substantial dietary source of antioxidant which may promote health and produce disease prevention effects.